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We used a novel phase-resolved optical Doppler tomographic (ODT) technique, with very high flow velocity sensitivity (10 μm/s) and high spatial resolution (10 μm), to image blood flow in port wine stain (PWS) birthmarks in human skin. The variance of blood flow velocity is used to locate the PWS vessels in addition to the regular ODT images. Our device combines an ODT system and laser so that PWS blood flow can be monitored in situ before and after treatment. To our knowledge, this is the first clinical application of ODT to provide a fast semi-quantitative evaluation of the efficacy of PWS laser therapy in situ and in real-time.

The glycosylation process is extremely heterogeneous, dynamic, and complex compared with any other post-translational modification of protein. In the context of recombinant glycoproteins, glycosylation is a critical attribute as glycans could dramatically alter protein functions and properties including activity, half-life, in vivo localization, stability, and, last but not least, immunogenicity. Liquid chromatography combined to mass spectrometry constitutes the most powerful analytical approach to achieve the comprehensive glycan profile description or comparison of glycoproteins. This chapter details a versatile yet straightforward LC-MS approach for sample preparation, analysis, and data interpretation, enabling the evaluation of site-specific N-glycosylation of recombinant glycoproteins.

Purified recombinant urate oxidase (urate oxygen oxidoreductase EC 1.7.3.3. re-Uox) has been studied by means of differential scanning calorimetry (DSC) in correlation with enzymatic activity measurements and size exclusion chromatography. Differential scanning calorimetry curves versus pH show two endothermal effects in the pH range 6-10. The first endotherm reveals a maximum stability between pH 7.25 and pH 9.5 corresponding to a temperature of transition T(m1) of 49.0 degrees C and an enthalpy of transition of 326 kJ mol(-1). This value dramatically decreases below pH 7.25. The behavior of the second endotherm is more complex but the temperature of transition T(m2) is constant between pH 9 and 7.25 and a maximum for the corresponding enthalpy is obtained near pH 8 with DeltaH(2)=272 kJ mol(-1). An optimal pH of 8.0 for the stability of the enzymatic activity at elevated temperature was also found which was in good agreement with calorimetric results. Reversibility of the first endotherm is obtained from 20 to 51.5 degrees C. The calorimetric result is correlated to enzymatic activity, purity by size exclusion chromatography (SEC) and protein concentration measurements. In contrast, for the second endotherm, after heating up to 68.9 degrees C, no reversibility was found. Interaction with structural analogues of urate has been studied by DSC. 8-Azahyooxanthine has only a small effect and caffeine has no effect at all. With 8-azaxanthine, a rapid increase of the T(m1) function of the concentration is obtained. At high concentration T(m1) reached the T(m2) value which remained unaffected.

The effect of manganese on lignin peroxidase activity was studied. The enzyme was produced with a new process using an air-lift-type reactor. The experiments were performed with veratryl alcohol and a dimeric lignin model compound. It was shown that when Mn(II).lactate complex was present the amount of veratraldehyde formed and the uptake of oxygen were significantly enhanced during the aerobic oxidation of veratryl alcohol. A similar effect can be obtained with superoxide dismutase. These results strongly suggest that the superoxide anion can occur during the reaction; its scavenging by Mn(II) or superoxide dismutase generates H2O2. In contrast, no evidence for the formation of superoxide anion was found during oxidation of the lignin model, compound veratrylglycerol-beta-guaiacyl ether.